16-keto steroids of the pregnane series and methods for making same



D of the molecule.

-- as hydrogen peroxide. appear from the detailed description of theinvention hereinbelow.

Unite 16-KETO STEROIDS F THE PREGNANE SERIES AND METHODS FOR MAKING SAMEDavid Taub, Metuchen, and Norman L. Wendler, Summit, N.J., assignors toMerck & Co., Inc., Rahway NJ., a corporation of New Jersey Thisinvention relates to a new class of oxygenated steroids. Moreparticularly, it is concerned with steroids of the pregnane serieshaving a ketone function in ring Still more particularly, it relates to16,20-diketo-17u-hydroxy steroids of the pregnane series, which newcompounds have the partial structural formula- It is, of course, wellknown that the pharmacological double bond in the 1:2 positions of the.steroid nucleus or by halogenation at the 9-position. The 'search'toimprove the activity of such hormones still further has been a difiicultone and several attempted modifications in the structure ofphysiologically active steroids have resulted in loss or diminution ofactivity. We have now discovered, however, that introduction of a ketonefunction at the 16-position of 17a-hydroxy-20-keto pregnanes gives riseto new compounds having increased anti-inflammatory activity without anysubstantial degree of undesirable side eifects, such as retention ofsodium.

It is therefore an object ofour invention to provide a new series ofsteroids of the prcgnane series, which compounds have a lc etone groupin the C-l6 position. It is a further object to provide l6,20-diketo17a-hydroxy steroids of the pregnane series and processes for makingsuch compounds from A -20-keto pregnenes. Still another object is asynthetic method of making 16,20-diketo- 17u-hydroxy steroids bytreatment of a A -20-keto steroid with a metal oxide and a peroxidecompound.

Such l6-keto steroids of the pregnane series are themselvesphysiologically active compounds having enhanced cortisone-like activityand of value in the treatment of inflammatory and allergic conditions,or are important intermediates in the synthesis of physiologicallyactive 16-keto-pregnenes or pregnadienes. A more particular object isthe synthesis-of 16,20-diketo-l7d-hydroxy steroids by reaction of a A-2O-keto'steroid with a metal oxide such as osmium tetroxide andaperoxide compound such Additional and further objects Will The newcompounds of our invention may be produced F Patent 70 "2 by treating aA -20 keto steroid with a metal oxide and a peroxide compound, as shownby the flowsheet:

The critical structural feature of the starting material for makingl6,20-diketo-l7u-hydroxy steroids of the pregnane series is the A-20-keto moiety. The molecule may be substituted or unsubstituted inother portions of the molecule and may also be unsaturated. Thus A A orA steroids, or. compounds having substituents such as keto, hydroxy,esterified hydroxy, chlorine, bromine, fluo rine and/or methyl groups onthe nuclear or 21-carbon atoms are suitable reactants. Double bonds inthe mole,- cule (other than the Al -bond) which are in hindered portionsof the molecule such as at the AW position or are conjugated with acarbonyl group, are inert under the conditions of our process. However,other double bonds are apt to react with osmium tetroxide-hydrogenperoxide and should be protectedichemically prior to the reaction inorder to avoid undesirable side reactions.

As the metal oxide in our process, we prefer to use osmium tetroxide,although other metal oxides which add to a carbon-carbon double bond,e.g. tungsten trioxide, may also be employed. The peroxide compoundwhich is preferred is hydrogen peroxide, other suitable peroxides beingalkyl peroxides such as t-butyl hy'droperoxide or peracids such asperacetic, perbenzoic or perphthalic acids. The oxidation is carried outa solvent inert under the reaction conditions. Suitable solvents areten;- tiary allcanols such as tertiary butanol and tertiary amylalcohol, halogenated hydrocarbons like carbon tetrachloride, ethylenedichloride, methylene chloride, ethyl ether, benzene, dioxane, ethylacetate and the like. i V

In this process, only relatively small percentages .of metal oxide,based on total steroid, are utilized. For optimum results we employabout 0.01 to 0.25 mole, and preferably about 0.1 mole, of osmiumtetroxide per mole of pregnane compound. However, an excess of peroxidecompound is desirable. Usually about a 100% excess, again based on thesteroid reactant, is used for best results, although from about to about200% excess is satisfactory.

To carry out the oxidation, one of the reagents is mixed with a solutionof the steroid reactant, and a solvent solution of the second reagentadded slowly thereto. We prefer to add an osmium tetroxide solutionslowly to a mixture of pregnane compound and hydrogen peroxide. However,the desired 16,20-diketo-l7a-hydroxy pregnane compound is produced ingood yield by the alternate method of adding the hydrogen peroxide tothe osmium tetroxide-steroid solution. In either case, the resultinglo-ketone may be isolated by the same standard tech-- niques.

At the preferred reaction temperatures of about 20-45* C. optimum yieldsof desired l6,20-diketo-l7a-hydroxy pregnane compounds are obtained inabout 1-43 hours, and preferably in 3-10 hours, although considerably 3longer reaction times do not appear to affect the reaction adversely andmay be utilized if desired.

At the completion of the reaction, the mixture is ordinarily treatedwith a small amount of reducing agent such as sodium sulfite or hydrogensulfide for a sort period of time, after which the desired16,20-diketo-17u-hydroxy steroid is recovered in substantially pure formby methods well known to those skilled in theart. Extraction into anorganic solvent and crystallization from that solvent or from a mixtureof solvents, either with or without chromatographic purification, aresatisfactory techniques for isolating our new steroids.

The above discussion of our processfor making 16,20- diketo-lh-hydroxysteroids of the pregnane series from A -20-keto steroids is notrestricted to specific members of the series since the oxidationreaction is of general application and does not depend for its successon the presence or absence of functional substituents or unsaturation inother portions of the molecule.- For optimum yields in our process, weprefer that steroids having a 2l-hydroxy group, be esterified at thatposition, preferably by formation of an ester with an organic carboxylicacid. Typical acids which may be used for this purpose are acetic,propionic, butyric, t-butylacetic, stearic, benzoic, oxalic, succinic,tricarballylic, phenylacetic, phthalic, dimethylacetic, trimethylaceticand the like. Likewise, the C-21 ester may be formed from an inorganicacid, for example sulfuric and phosphoric acids. In these cases, thecorresponding 16,20-diketo-17a-hydroxy-pregnane 21- -ester is produced.

Of particular importance are 16,20-diketo-17a-hydroxy steroidsunsaturated in ring A in the 1:2 and/or 4:5

9u-fluoro-A -pregnadiene-llfl,l7u,21 triol 3,16,20-trione 2l-propionate;9a-fluoro-A -pregnadiene-11fl,17u,21-triol 3,16,20 trione2l-cyclopentylpropionate; 9u-fluoro-A -pregnadien-l7a,2l-diol-3,11,16,20tetrone ZI-acetate; A -pregnen-l7a,2l-diol-3,l1,16,20-tetrone21-butyrate; M-pregnen-l7u,21-diol-3,11,16,20-tetrone 2l-acetate; A-pregnene-3,16,20-trione; A -pregnen-17a,21-diol-3,16,20-trione2l-acetate; n. -pregnen-3,17a,21-triol-l6,20-dione-3,2l-diacetate; and9a-chloro-A -pregnene-11B,17o ,21 triol 3,16,20-trione 21aceta-t'e.

1 It will be appreciated by those experienced in this .art that, insynthesizing 16,20-diketo-l7a-hydroxy pregnanes functionally substitutedor unsaturated in other portions positions and having the functionalsubstituents shown in formula I:

of the molecule, the 16-keto17a-hydroxy functional moiety may beintroduced in the final step of the synthesis from the appropriate A-compound, or it may be integrated into the molecule at an earlierstage, and other desired functional groups such as 1:2 and/or 4:5 doublebonds, ll-oxygenated groups, 9 -halo substituents, 2 l-hydroxyor21-ester functions introduced by known reactions into the16-keto-17u-hydroxy compound. For instance, in synthesizing thephysiologically active 9a-fluoro-A -pregnadiene-l13,17a,2l-triol-3,16,20-trione 21-acetate, one may employ 9ot-fiuoro-A-pregnatriene 116,21-diol-3, 20-dione 2l-acetate as an intermediate inwhich case the desired compound is obtained directly by the hereindisclosed process. Alternatively, when the osmium tetroxide-hydrogenperoxide reaction is carried out on 9a-fluoro- A-pregnadiene-11fi,2l-diol-3,20-dione 21-acetate or on A-pregnadiene-I1,8,21-dio1-3,20-dione 21-acetate, the desired compoundwill not be obtained directly but can be readily made from the16-keto-17a-hydroxylated prod not by introduction of a 1:2 double bondas described in zoic, oxalic,-or phenylacetic and inorganic acids suchas sulfuric andphosphoric may be used to form the ester. These16,20-diketo-17whydroxy steroids, which are obtained from thecorresponding A -20-keto steroid compounds have pronouncedcortisone-like activity.

16,20-diketo-17a hydroxy steroids of the pregnane series which may bemade by the process of this invention from A -20-keto steroids arehours.

trioxide, but other such as sodium dichromate, N-bromsuccinimide andN-bromacetamide may be employed. In this process, the oxidizing agentwill act on other oxidizable groups in the molecule as well as on the '16-hydroxy group; so that -suchother functions should be suitably blockedor protected, as by esterification, prior to the oxidation. Using thisprocess, we have, for instance, prepared A '-pregnen-l7u,2l di0l-3,l1,16,20-tetrone 21-acetate l6-keto cortisone acetate) from A-pregnen-16u,l7oi,

21-triol-3,11,20-trione 21-acetate (16,- hydroxycortisone acetate). Thechromium trioxide oxidation is preferably efiected in a suitably inertsolvent medium such as acetic acid or pyridine at about 20 C. to about35 C. At these temperatures optimum yields of 16-ketones are realized inabout 5-20 hours and usually in about 5-15 At increased temperatures,the reaction times may, of course, be reduced.

Our new physiologically active 16,20-dik8t0-17oc-1IY- droxy steroids maybe administered orally as tablets or capsules as anti-inflammatoryagents or applied topically in the form of lotions or creams. A suitabletablet is prepared having the following ingredients:

Magnesium stear 0.002

tion, washed with water and dried.

The steroid, lactose, corn starch and sucrose are mixed and wet with theacacia. The mass is then forced through a No. 8 mesh screen and dried,and the dried granules forced through a No. 12 mesh screen.- Thesegranules are mixed with the magnesium stearate and compressed intotablets using a fraction inch deep cup punch. If desired, they may becoated with sugar by standard techniques.

A topical ointment containing one of our new physiologically activel6-keto steroidsmay be prepared by mixing 0.025 gram of steroid withabout 0.1 gram of propylene glycol, milling the resulting paste andadding an additional 0.2 gram. of propylene glycol. This mixture is thenadded to about 042 gram ofmilled polyethylene glycol, about 0.5 ml. ofWater is then added, the mixture stirred and powdered zinc stearatemixed in until the mass is partially congealed. The ointment is thenmilled.

Certain-of the l6,20 -diketo-17a-hydroxy pregnane compounds-embracedwithin our invention are useful primarily as intermediates in makinghighly active 16-keto compounds. They are so converted by theintroduction, by known methods, of functional moieties such as a. 1:2double bond, a 4:5 double bond, at 9-halo substituent, an

ll-oxygenated function, a 2l-hydroxy group or a 3-keto group into highlyactive 16,20-diketo-17a-hydroxy steroids of the pregnane series.

The following examples are given for purposes of illustration and not byway of limitation:

EXAMPLE 1 i 9a-fluoro-A -pregnadiene-11,8,21 -diol-3,20-dione 21-acetate *20-dione 2l-acetate is dissolved in ml. of methanol and ofsemicarbazide hydrochloride and 0.74 gram of sodium bicarbonate in 3 ml..of water. The reaction mixture is stirred at reflux temperature forthree and one-half hours and then at -45" C. for 16 hours. The resulting3,20- diser'nicarbazone derivative of the. starting material isprecipitated, by cooling and addition of 200 ml. of saturated. sodiumchloride "solution. It is isolated by filtra- One gram of the3,20-disemicarbazone obtained above and-'20 of acetic acid and 1 ml. ofacetic anhydride .are iefluxed under a nitrogen atmospherefor one hour.

There is thus obtained a solution of the 3,20-disemicarbazone of9a-fluoro-A -pregnadiene 11fl,2l diol-3,20- dione 2l-acetate. Withoutisolation of this material, the reaction mixture is concentrated invacuo to 10 ml. and

treated with 5 ml. of water and 3 ml. of pyruvic acid.

The mixture is allowed to stand at room temperature for about'24 hours,after which time water is added and the mixture extracted withchloroform. The chloroform extract is washed with Water and potassiumbicarbonate solution, and dried over magnesium sulfate. The chloroformsolution is chromatographed on about 25 grams of neutral alumina and9ot-flu0ro-A -pregnadiene-1113,21- diol-3,20-dione 21-acetate obtainedby concentration of the benzene-chloroform eluted fractions.

EXAMPLE 2 Qa-fluOr -M -pregnadi en-21 -0l-3,1 1 ,20-lri0ne 21 -acemtewater. The reaction mixture is heated at reflux for three hours and at40 C. for ten hours. The resulting 3,20- disemicarbazone derivative ofthe starting material is 3 precipitated by addition of water and coolingin an ice bath. It is isolated by filtration and dried.

The disemicarbazone obtained above is refluxed for one hour in an inertatmosphere with 10 ml, of acetic acid and 1 ml. of acetic anhydride. Thereaction mixture is then concentrated to about one-half the volume andtreated with water and 2 ml. of pyruvic acid. At the end of 15 hours at25 C., additional water is added and the mixture extracted with benzene.The benzene extract is washed with water and sodium bicarbonatesolution, and dried. On chromatography over 12 grams of neutral aluminathere is obtained in the benzene eluted fractions 9a-fluoro-A-pregnadien-2 l'ol-3, 1 1 ,ZO-trione 2 l acetate.

I EXAMPLE 3 M-pregnene-I 1 6,1 7a,21 -tri0l-3,1 6,20-frione 21 acetateTo a stirred solution of 116 mg. of d pregnadiene-1l,3,21-diol-3,20-dione 21-acetate and 3 ml. of t-butanol is added 1.63ml. of 0.37 M hydrogen peroxide in t-butanol and 8 mg. of osmiumtetroxide in 1 ml. of 't-butanol. The addition is carried out dropwise.The reaction is allowed to proceed for 48 hours at room temperature,after which time Water is added and the mixture extracted withchloroform. The chloroform extract is concentrated in vacuo and theresidue dissolve in 10 ml. of methanol. mg. of sodium sulfite in 2 ml.of water is added to the methanolic solution and the mixture refluxedfor 30 minutes. After cooling and addition of more water the mixture isextracted with chloroform. The chloroform is again removedbydistillation and the residue treated at room temperature for fivehours with 0.5 ml. of acetic anhydride in 1 ml. of pyridine. Ten ml. ofWater is then added to the back-acetylation reaction mixture and thedesired steroid extracted with chloroform. The chloroform extract ischromatographed on about three grams of neutral alumina. Elution of thecolumn with benzene-chloroform and concentration of the resultingeluates yields A -preg- 116118 1lB,17oz,2it1'i0l-3,16,Z0-tii0l16ZI-acetate.

The starting material for this reaction may be prepared as described inthe Journal of the American Chemical Society 77, 1028 (1955).

EXAMPLE 4 A -pregnen'-17a,21 dial-3,1 1,1 6,20-tetr0-ne 21 -acetate To asolution of 200 mg. of A -pregnadien-2l-ol-3,- l-LZO-trione 21-acet atein 10 ml. t-butanol is added five drops of pyridine and 5 mg. of osmiumtetroxide in 5 ml. t-butanol. Five ml. of a 0.4 M solution hydrogenperoxide in t-butanol is then added slowly over a period of 45 minutes.The resulting mixture is then stirred for an additional 30 minutes atroom temperature. At the end of this time 300 mg. of sodium sulfite in15 ml. of water is added and the resulting mixture stirred for about 10minutes. It is then concentrated to about one-third the volume in vacuoand extracted with chloroform. The chloroform extract is washed withWater and dried, and then chromatographed over 5 grams of neutralalumina. On elution of the column with benzene-chloroform, A pregnenl7a,21 diol 3,11,16,20 tetrone 21- acetate is found in the eluate andmay be isolated by concentrating in vacuo.

EXAMPLE 5 Ai-pregnene-J 1 8,1 7a,.21-tri0Z-3,16,2O-trione To a stirredsolution of 100 mg. of A-pregnene-llfl;

l7a,21-triol-3,16,20-trione 2lacetate in 40 ml. of 90% methanol isadded, at 20 C., 2 ml. of concentrated hydrochloric acid. The mixture isallowed to stand at about 25 C. for six hours and then ml. of water isadded. The resulting suspension is cooled and the crystalline purities.

A? pregnene 11;.3,17a,21 triol 3,16,20 trione filtered, washed withwater and dried in vacuo.

, EXAMPLE 6 v3 ml. oft-butanol is added 1.63 ml. of 0.37 M hydrogenperoxide in t-butanol. Eight mg. of osmium tetroxide in .1 ml. oft-butanol is added to the steroid dropwise over a period of about 30minutes. The reaction mixture is allowed to stand at room temperaturefor 38 hours and, at the end of this time water is added and the steroidextracted with chloroform as described in Example 3. Following theisolation procedure set forth in Example 3 .there is obtained9a-fiuoro-A -pregnene-11,B,17a,21-trio1- -3,16,20-trione 21-t-butylacetate.

EXAMPLE 7 A -pregnadiene-1IB,1 7 04,21 -triol-3,I 6,20-trione, an21-acetate I Fifty ml. of a nutrient medium are prepared having thefollowing composition:

'Cerelose grams 1 Edamin do. 1 Cornsteep liquor ml 0.25

Distilled water to make 50 ml.

This medium is adjusted to pH 6.5 with potassium hydroxide, sterilizedand inoculated with about 2.5 to ml. of a culture of Bacilfus sphaericus(MB 431) microorganisms, and the inoculated culture is then incubated ata temperature of 28 C., with agitation, for a 24-hour period.

To the resulting culture is added a solution containing 10 mg. of A pregnene-l1,8,17ot,21-triol-3,l6,20-trione 21- acetate dissolved in 0.2 ml.of dimethylformamide. The culture containing the steroid compound isincubated, with agitation, for an additional period of about 24 hours at28 C.

The fermentation broth is extracted with four 50 m1.- portions of ethylacetate, and the ethyl acetate extracts are combined and evaporated todryness in vacuo. The

residual material is dissolved in acetone and treated with charcoal anddiatomaceous earth to remove colored im- The decolorized acetonesolution is then streaked on paper chromatograms which are developedusing formamide as the stationary phase and chloroform as the mobilephase. The upper bands are removed, extracted with methanol and themethanol extract again chromatographed by the paper streak method. Theupper band is cut off, dried, extracted with methanol and the methanolextract concentrated to dryness in vacuo.

.7 small amount of pyridine.

Fifty ml. of a nutrient medium are'p'repared having the followingcomposition:

Cerelose gr ams .l Edamin d0 l Cornsteep liquor ml 0.25

Distilled Water to make 50ml.

This medium is adjusted to pH 6.5 with potassium hydroxide, sterilizedand inoculated with about 2.5 to 5 ml. of a culture of Bacillussphaericus (MB 431) microorganisms, and the inoculated culture is thenincubated at a temperature of 28 C., with agitation, for a '24-hourperiod. To the resulting culture is added a solution con taining 10 mg.of L-fll10IO-A -P1'egl'1fl'1-17,21-dlOl-3,11',

16,20-tetrone ZI-acetate dissolved in 0.2 ml. of dimethyl- EXAMPLE 99a-flu0r0-A -pregnadiene-I113,17a,21-tri0l-3,I6,20-tri0ne ml; of anutrient medium are prepared having'the following composition:

Cerelose ams" 2 Edamin 4 7 do 2 Cornsteep liquor Distilled water to make100 m1.

This medium is adjusted to pH 6.5 with potassium hydroxide, sterilizedand inoculated with about 5 to 10 ml. of a culture of Bacillussphaericus (MB 431) microorganisms, and the inoculated culture is thenincubated at a temperature of 28 C., with agitation, for a 24-hourperiod. To the resulting culture is added a solution containing 20 mg.of 9a-fiuoro-A pregnene-l1fi,17a,21-triol- 3,16,20-trione dissolved in0.4 ml. of dimethylformamide. The culture containing the steroidcompound is incubated, with agitation, for an additional period of about24 hours at 28 C.

The fermentation broth is extracted with four 100 mL-portions of ethylacetate, and the ethyl acetate extracts are combined and evaporated todryness in vacuo. The residual material is dissolved in acetone andtreated with charcoal and diatomaceous earth to remove coloredimpurities. The decolorized acetone solution is then streaked on paperchromatograms which are developed using formamide as the stationaryphase and chloroform as the mobile phase. The upper bands are removed,extracted with methanol and the methanol extract again chromatographedby the paper streak method. The upper band is cut off, dried, extractedwith methanol and the methanol extract concentrated to dryness in vacuo.The residue is crystallized from a mixture of ethyl acetatepetroleumether to give crystalline 9oc-fiuoro-A-pregnadiene-l1fi,17a,21-triolr3,16,20-trione. p a

The corresponding 21-acetate derivative of the above product is obtainedby refluxing the steroid for 45 minutes with 6 ml. of acetic acid and0.5 ml. of acetic anhydride. The hot reaction mixture is then cooledandconcentrated to a small volume and treated with 5 ml. of water. The9a-iluoro-A -pregnadieri-11fl,17a,21 triol- 3,16,20-trione' 21-acetateis isolated by extracting the mixture with chloroform and removal of thechloroform V by distillation.

M-pregnen-I 7a,21-di0l-3,11,1 6 ,2-t'etr0ne 21 acetate A solution of 105mg. of A -pregnen-16u,l7u,2l-triol- 3,11,20-tri0ne 21-acetate [preparedby the method described in J. Chem. Soc. 4383 (1955)] in 2 ml. of aceticacid is treated with 16.7 mg. of chromium trioxide in 0.1

ml. acetic acid. The reaction mixture is allowed to stand at 25 C. for16 hours, after which water is added and the steroid is extracted withchloroform. Paper streak chromatography using a benzene-forrnamidesystem shows that the main product of the reaction is A -pregnen-17u,21-diol-3,11,16,20-tetrone 21-acetate. Substantially pure materialis obtained by chromatography of the chloroform extract. over neutralalumina. The desired 16-'ketone is isolated from the benzene-chloroformeluates and has a melting point of 178-180 C.

EXAMPLE 11 8 ml. of 0.2 M hydrogen peroxide in t-amyl alcohol is addedslowly to a mixture of 210 mg. of 9a-fluoro- A-pregnatriene-11/3,21-diol-3,20-dione 21-acetate and 14 mg. of osmiumtetroxide in 10 ml. of t-amyl alcohol.

The reaction mixture is stirredat about 30 C. for ten,

hours. At the end of this time water is added and the mixture extractedwith two 10 ml. portions of chloroform.

The chloroform is removed bydistillation and the residual materialdissolved in 15 ml. of methanol. Excess hytion for 15 minutes. extractedwith fresh chloroform.

The chloroform extracts are washed with water, dried over magnesiumsulfate and poured over grams of dr'o'g'en sulfide gas is bubbledthrough the methanol solu- Water is added and the mixture is alumina.Elutiorl of the alumina with benzene-chloroform, and removal of theorganic solvents in vacuo gives substantially pure 9a fluoro-A-pregnadiene-I1B,17a-21- triol-3,16,20-trioi1e ZI-acetate.

The 90c fluoro A1346 pregnatriene 1119,21 diol- 3,20 dione 21 acetateused in this experiment is prepared from 90: fluoro A pregnadiene115,17a,21- triol 3,20 dione 21 acetate by formation of a 3,20- bissernicarbazone derivative, dehydration with acetic acid and reversal ofthe semicarbazone as described in Example 1.

In a similar manner 90: chloro A pregnadiene- 11fi,17a,21 triol 3,16,2 0trione 21 benzoate; 9a-

nuern A pregnadien 17a,2l diol 3,11,16,20- tetrone and 9a fluoro Apregnadiene 115,17- diol 3,16,20 trione are prepared by treatment of thecorresponding A compounds with osmium tetroxide and hydrogen peroxide.

EXAMPLE 12 12a fluoro A pregnadiene 11fl,]7a,21 triol 3, 16,20 trione21. acetate 2 ml. of pyridine and 7 mg. of osmium tetroxide in 7 m1. oft amyl alcohol are added to a solution of 300 mg. of 12a fluoro Apregnadiene 11,8,21 diol- 3,20 dione 21 acetate in 15 ml. of t amylalcohol. To this mixture 10 ml. of a 0.035 M solution of hydro genperoxide in t amyl alcohol is added dropwise over two hours. Thereaction mixture is then stirred at 25 C. for an additional 40 minutes,and then hydrogen.- sulfide gas bubbled through the solution for 20minutes.

The mixture is then extracted with two 10 ml. portions of chloroform,the chloroform extracts combined, washed with water and dried. Theextract is concentrated in vacuo to a small volume and chromatographedover seven grams of neutral alumina. The column is eluted withbenzene-chloroform andthe eluates concentrated to dryness, giving aresidue of 12a fluoi'o A pregnadiene- 11,3, 170:, 21 triol 3,16,20trione 21 acetate.

In a similar fashion 12a chloro A pregnen- 17a, 21 diol 3,11,16,20tetrone 21 acetate; 12o: fluoro- A pregnene 11 3,l7oz,2l triol 3,16,20trione 21 t butyl acetate and 12a fluoro A pregnadien 17a, 21 diol3,11,16,20 tetrone are obtained from the corresponding A compounds. Inthe case of the free 21- alcohols, it is preferable to carry out theoxidation on a 21 acetate, and subsequently remove the ester function bythe method of Example 5. The starting A steroids are prepared by theprocedure of Example 1 from the corresponding 17o: hydroxy steroids.These latter compounds are made by the processes disclosed and claimedin our copending application Serial No. 590,828, filed June 12, 1956,now abandoned and by the methods described in Chemistry and Industry,October 27, 1956, page 1232.

Any departure from the above description which conforms to thepresentinvention is intended to be included within the scope of the claims. .1

What is claimed is:

1. A pregnene 11fi,17a,21 triol 3,16,20 trione.

2. A pregnene 11,8,17a,21 triol 3,16,20 trione 21 acetate. 1

3. A pre'gnene 11,8,17a,21 triol 3,16,20 trione 2'1 tertiary butylacetate.

4. 9a fluoro A pregnene 1lf3,l7a,21 triol 3,

16,20-:trione. a

5. 9oz fluoro A pregnene 1lfl,17vt,2l -.'triol 3, 16,20 trione 21acetate. 7 i

6. A pregnadien 17a,2l diol 3,11,16,20 tetrone 21 acetate. 1

'7. A pregnadiene 1l,8,l7ot,21 triol 3,16,20 trione. a

8. A pregnadiene l1;8,17a,21 triol 3,16,20 trione 21 acetate.

9. 9a fluoro A pregnadiene llfi,l7a,2l triol- 3,16,20 trione.

10. c fiuoro A pregnadiene 11B,17u,2l triol- 3,16,20 trione 21 acetate.

11. 9oz fluoro A pregnadiene l1B,17o:,21 triol- 3,16,20 trione 21 tbutyl acetate.

12. The process which comprises treating 3,11 bisoxygenated A 20 ketosteroids of the pregnane series with osmium tetroxide in the presence ofa peroxide compound, thereby producing 3,11 bis oxygenated -16,20-diketo 17a hydroxy steroids of the pregnane series, wherein theoxygenated functions at positions 3 and 11 are selected from the classconsisting of hydroxy and keto groups.

13. The process which comprises treating 3,11 bisoxygenated A 20 ketosteroids of the pregnane series with a catalytic amount of osmiumtetroxide in the presence of a peroxide compound, thereby producing3,11- bis oxygenated 16,20 diketo A 17oz hydroxy steroids of thepregnane series, wherein the oxygenated functions at positions 3 and 11are selected from the class consisting of hydroxy and keto groups.

14. The process which comprises treating 3,11 bisoxygenated A 20 keto.steroids of the pregnane series with a catalytic amount of osmiumtetroxide in the presence of hydrogen peroxide, thereby producing 3,11-bisoxygenated 16,20 diketo 17a hydroxy steroidsof the pregnane series,wherein the oxygenated function-s at p0- sitions 3 and 11 are selectedfrom the class consisting of hydroxy and keto groups.

15. The process which comprises treating A4116 pregnadiene 1119,21 diol3,20 dione 21 acetate with osmium tetroxide and hydrogen peroxidethereby producing A pregnene 11fl,17 x,21 triol 3,16,20 trione 21acetate.

16. The process which comprises treating 90c fluoro- A pregnadiene 118,21 diol 3,20 dione 21 acetate with osmium tetroxide and hydrogenperoxide thereby producing 904 fluoro A pregnene 11,B,170t,21- triol3,16,2- trione 21 acetate.

17. The process which comprises treating 90c fluoro- A pregnadiene115,21 diol 3,20 dione 21 tbutyl acetate with osmium tetroxide andhydrogen peroxide thereby producing 90: fiuoro A pregnene 11152, 1741,21triol 3,16,20 trione 21 t butyl acetate.

18. The process which comprises treating A1416 pregnatriene 115,21 diol3,20 dione 21 acetate with osmium tetroxide and hydrogen peroxidethereby producing A pregnadiene 11,8,17a,21 triol 3,16,20 trione 21acetate.

19. The process comprising treating 3,20 diketo 11- oxygenated 16,17adihydroxy pregnanes having a double bond attached to the C- carbon atomwith chromium trioxide thereby producing 3,16,20 triketo- 11 oxygenated170a hydroxy pregnanes having a double bond attached to the C-5 carbonatom, wherein the 11- oxygenated function is selected from the classconsisting of hydroxy and keto groups.

20. The process which comprises treating 9a fluoro- A pregnatriene115,21 diol 3,20 dione 21- acetate with osmium tetroxide and hydrogenperoxide thereby producing 9a fluoro A pregnadiene llfl, 17a,21 triol3,16,20 trione 21 acetate.

21. 3,16,20 triketo 11 oxygenated 17a hydroxy steroids of the pregnaneseries having a double bond in the 4:5 position, wherein the oxygenatedgroup at the 11 position of the steroid nucleus is a hydroxy group.

22. A member of the class consisting of steroids having the formulawherein X is selected from the class consisting of keto and hydroxygroups, Y is selected from the class consisting of hydrogen and halogen,and 0-21 lower alkanoates thereof.

12 24. A compound having the formula 15 wherein R is the residue of alower aliphatic acid.

25. A compound having the formula OHaOR onion l wherein'R is the residueof a lower aliphatic acid and X 5 is halogen. V r

27. A compound having the formula OHaOR- C.=O

"'OH HO =0 /r 13 14 with osmium tetroxide and hydrogen peroxide therebywith osmium tetroxide and hydrogen peroxide thereby forming a compoundhaving the formula forming a compound having the formula 132011 0111012:0 (1:0 -"OH ----0H xfi: To Xfj To l Y Y wherein X is selected from theclass consisting of keto wherein X is iselqcted from class wnsisting..fketo and hydroxy, Y is selected from the class consisting of a hydroxy Y18 selected m the class conslstmg of hydrogen and halogen, and R isselected from the class P F and halogen and R 15 Selected P the classconsisting of hydrogen and lower acy1radic 1s consistlng of hydrogen andlower acyl radicals.

29. The process which comprises treating a compound having the formulaReferences Cited 1n the file of th1s patent 20 UNITED STATES PATENTS E 22,901,492 Herz et a1. Aug. 25, 1959

1. $4-PREGNENE - 11B,17A,21 - TRIOL - 3,16,20 - TRIONE.